Isolation of RNA from Field-Grown Jute (Corchorus capsularis) Plant in Different Developmental Stages for Effective Downstream Molecular Analysis

• Published in: Molecular biotechnology 2011-10, Vol.49 (2), p.109-115
• Main Authors: Samanta, Pradipta, Sadhukhan, Sanjoy, Das, Subrata, Joshi, Alpana, Sen, Soumitra K, Basu, Asitava
• Format: Article
• Language: English
• Subjects: Agricultural biotechnology; Biochemistry; Biological and medical sciences; Biological Techniques; Biotechnology; cDNA libraries; Cell Biology; Centrifugation; Chemistry; Chemistry and Materials Science; complementary DNA; Corchorus - genetics; Corchorus - growth & development; Corchorus capsularis; crops; Developmental stages; Economic importance; Electrophoresis, Agar Gel; Fundamental and applied biological sciences. Psychology; Gene expression; Gene Library; Genetic engineering; genetic improvement; Human Genetics; jute; Molecular biology; phenolic compounds; Phenols; Plant biology; Plant species; Plant Stems - chemistry; Plant Stems - growth & development; Plant tissues; Protein Science; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic acid; RNA; RNA, Plant - analysis; RNA, Plant - isolation & purification; Textiles
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-011-9376-8

Jute (Corchorus capsularis), as a natural fibre producing plant species, ranks next to cotton only. Today, biotechnological approach has been considered as most accepted means for any genetic improvement of plant species. However, genetic control of the fibre development in jute has not yet been explored sufficiently for desired genetic improvement. One of the major impediments in exploring the genetic architecture in this crop at molecular level is the availability of good quality RNA from field-grown plant tissues mostly due to the presence of high amount of mucilage and phenolics. Development of a suitable RNA isolation method is becoming essential for deciphering developmental stage-specific gene expression pattern related to fibre formation in this crop species. A combination of modified hot borate buffer followed by isopycnic centrifugation (termed as HBIC) was adopted and found to be the best isolation method yielding sufficient quantity (~350–500 μg/gm fresh tissue) and good quality (A260/280 ratio 1.88 to 1.91) RNA depending on the developmental stage of stem tissue from field-grown jute plant. The poly A+ RNA purified from total RNA isolated by the present method was found amenable to efficient RT-PCR and cDNA library construction. The present development of RNA isolation was found to be appropriate for gene expression analysis related to fibre formation in this economically important jute plant in near future.